Apparatus for making fluorophotometric measurements



H. L. coNmg F APPARATUS FOR MAKING nu OPHOTOMETRIC MEASUREMENTS Filed July 8, 1948 1 N V EN TOR. //0W/7ED L k. Ca/vo/FF Patented Apr. 17, 1951 UNITED STATES rm'rsnr ormcs;

I METRIC MEASUREMENTS Y Application July 8, 1948, Serial No. 37,566

4 Claims.

This invention :relates to an improved appa-w ratus for making fiuorometric measurements and-more particularly to improvements in cells for containing the sample ..used in making such measurements. The most accurate methods for ment cell or cuvette of Figure 5. assaying materials for content of :certain .vita- Throughout the drawings correspondingvnumins, notably thiamin andriboflavin, are almost merals refer to the same parts. 1 I 1 prohibitively tedious and time-consuming, and Referring to the drawingspFigure 1, there :is' methods have therefore been devised utilizing shown a light source lfl-provided'with a reflector the principal of -fiuorescence. In accordance i! for concentrating th'elight-ofthe light "source with 'such prior methods", ultra-violet light is proupon a lens 12, the light source, mirror and lens jected into a'sample of the substance undergoing being supported by the tube-13 which also serves examination, and the light} causing fluorescence to support one or-morefilters, when used; The in the sample, is then measuredand the degree light beam, which is determined by the filter, is of fluorescence"determined. The degree of fluo- 15 shown at 15 and isdirectedagainst one "face I! rescence is indicative of the content of riboflavin of a fiuorometric measurement cell' generally and certain other factors of the sample, and, designated l6. therefore, by comparing the fluorescence read- The face l1of thefiuorometricmeasurement ing with that of standardized samples, it is poscell, sometimes called a cuvette is provided sible in a-very short time to determine the apwith a window Z-B-and the face I8-ofthe cell proximate thiaminand riboflavin contents of is provided with a window 2|. The exte'riorsurvarious materials. face of the cell [6 is otherwise completely coated It is an object ofthepresent invention to imwith a mirror coating shown stippled in-the=-' prove the apparatusheretoforeavailable for such drawings. The coatingserves to provide a re fiuorometric measurements and to provide appafiective surface-which is reflective-toward theinratus in which the degreeof fluorescence and terior of the celllB throughout the entire cell photoelectric response of such fluorescence is wall area except for the window ZB aIIdthe wingreatlyincreased as compared with that obtained dow 2|. by prior apparatus. The photocell 20 is positioned-so as toviewthe It is a further object of the invention to prosample through the window-2l and Where desired vide an improved container-for-samples used in a fllter 19-' may be positioned'between the photomaking fiuorometric measurements and more cell and the face [8 of the fiuorometric cell. 'The' particularlyto' provi'de-wa 'cuvette or sampleconphotocell Zilis connectedbyleadwire's 22 to a"; tainer which is provided with a reflective interior suitable measuring -instrument--23-of known decoating throughout the entire exterior area of sign. I the sample container save only for the portions Referrin to F u 2 a d 3, the l ht beam through which excitation light is projected into [5 shines throu the W d W 25 to he nthe container and through which the fluorescent terior of the cell. The window 2! is, in accordresponse is observed photoelectrically. ance with the present invention, positioned so Other and further objects of the invention are 40 that its aX S is a a a le to the axis of the those inherent in the apparatus herein illuswindow 26 so that light shinin to the cell trated, described and claimed. through window 26 does not shine out of the The invention is illustrated with reference to cell through the window 2|. Accordingly, the the drawings in which only light which is visible through the window 2! Figure 1 is a schematic side elevational view is that due to the fiuoroescence of the material of the apparatus; within the cell.

Figure 2 is an isometric view showing certain Fluorescence is light radiation, presumably of the part-s separated for better understanding; caused by the migration of electrons from one Figure 3 is a plan view of the fiuorometric shell to anotherwithin the atom of a substance measurement cell, filter and photocell portion of when certain wave lengths of light strike the the app us f Figure l molecule and hence it is desirable that the photo- Figure 4 is another isometric view of the samcell pick up the fluorescent light only. Thus, the ple container only of the'apparatus of Figures 1, light beam [5 shiningthrough the window 26 into d the sample S in the cell causes fluorescence. of Figure 5 is a plan view of a modified form of the sample and the lightv beams denoted by 27 l the invention utilizinga circular or cylindrical fiuorometric measurement cell or cuvette; 7

Figures 6 and 7 are side and front elevational views, respectively,- of the fiuorometric measurearrows 21, Figure 3, which are indicative of the fluorescence, shine out against the photocell 20 and produce a photoelectric response thereon. Where used the filter I9 is positioned between the photocell 20 and the window 2| in the face l8 of the fluorometric cell It.

In accordance with the present invention the fiuorometric cell may also be of tubular design, thus in the shape of a test tube as illustrated in Figures 5, 6 and 7. In this form of the invention the entire exterior surface of the tube 3|] is covered with a mirror coating and through the mirror coating there is provided at least one window 3| through which an excitation light beam may be projected into the sample Within the cell, and one or more windows 32 which are positioned at an angle and in relation to window 3| such that light entering the cell through the window 3| does not directly shine out through the window 32. In this modified form of the invention, as in respect to that previously described, there is provided a photocell 35, Figure 5, which is connected by leads 3! to a suitable indicating apparatus. Where desired a filter 36 may be provided between the photocell 35 and the fluorometric cell 30.

In accordance with the present invention it is possible to obtain photoelectric response'which is about 300% or more as compared with that obtained when the sample is contained in a clear glass cuvette of identical shape and size. Thus, by providing a mirrored surface on the outside of the cuvette in which the sample is contained and by providing an aperture through the mirror coating for projecting a light beam into the interior of the cuvette and another appropriately placed for viewing the interior of the cell for making the photoelectric measurement therethrough, it is possible to obtain a very much greater photoelectric response than ever before possible. This is of especial advantage since it has been proven that small sample sizes give higher values in assaying most substances for vitamins, particularly thiamin and riboflavin, and therefore by utilizing the present apparatus, the sample size with which the analyst can work accurately is reduced to approximately one-third of the size previously used.

As many apparently widely different embodiments of this invention may be made without departing from the spirit and scope thereof, it is to be understood that I do not limit myself to the specific embodiments herein, except as de-. fined by the appended claims,

What I claim is:

1. An improved fluorometric cell comprising a light transparent container having spaced oppositely disposed walls of transparent material, a mirror coating on the exterior of said walls, one light window through said coating for allowing a light beam to be directed into the cell and a viewing window through said coating out of alignment with the first window through which photoelectric measurements can be taken of the light conditions within the cell.

2. The improved fluorometric cell of claim 1 further characterized in that the cell is shaped like a test tube, and the windows through the mirror coating thereon are positioned so that their axes are at an angle to each other.

3. An improved fiuorometric device comprising a fluid tight cell of rectangular cross-sectional shape having sides and a bottom, the cell walls being of light transparent material having a mirror coating on the outside so as to present a highly reflective mirror surface toward the interior of the cell, a light transmitting window through one of the container walls so as to permit light to be directed into the interior of the cell and another window through another cell wall and out of alignment with the first window so as to allow fluorometric measurements to be taken of the light conditions within the cell.

4. The improved fiuorometric cell of claim 3 further characterized in that the cell is composed of glass walls mirrored on the outside.

HOWARD LEE CONDIFF.

REFERENCES CITED The following references are of record in the file of this patent:

UNITED STATES PATENTS FOREIGN PATENTS Country Date France Apr. 15, 1935 Germany May 20, 1939 Number Number 

